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Hi everybody. I have a problem: I am trying to set up a typical NADH coupled assay to measure Pyrvate formation (using LDHase). One of the proteins of my system is a transmembrane protein, and I have to use it as a membrane extract. The problem is that in the membrane there seems to exist a second protein which interferes with the coupled assay, because it consumes NADH by a different and unknown pathway (I have not found yet the substrate being reduced). The solution would be very simple if I can find any general irreversible inhibitor for proteins which use NADH.
Does anybody know any such inhibitor??
Thank you very much for any information.
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<DIV><FONT color=#000000 face=Arial size=2>&nbsp;&nbsp;&nbsp; <FONT
color=#000000>Hi everybody. I have a problem:&nbsp; I am&nbsp; trying to set up
a typical NADH coupled assay to measure Pyrvate formation (using LDHase). One of
the proteins of my system is a transmembrane protein, and I have to use it as a
membrane extract. The problem is that in the membrane there seems to exist a
second protein which interferes with the coupled assay, because it consumes NADH
by a different and unknown pathway (I have not found yet the substrate being
reduced). The solution would be very simple if I can find any general
irreversible inhibitor for proteins which use NADH.</FONT></FONT></DIV>
<DIV><FONT color=#000000 face=Arial size=2><FONT
color=#000000>&nbsp;&nbsp;&nbsp; Does anybody know any such
inhibitor??</FONT></FONT></DIV>
<DIV><FONT color=#000000 face=Arial size=2><FONT
color=#000000></FONT></FONT>&nbsp;</DIV>
<DIV><FONT color=#000000 face=Arial size=2><FONT
color=#000000>&nbsp;&nbsp;&nbsp; Thank you very much for any
information.</FONT></FONT></DIV>
<DIV><FONT color=#000000 face=Arial size=2><FONT
color=#000000></FONT></FONT>&nbsp;</DIV></BODY></HTML>
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